Dentin was visualized using a new fluorescence technique and confocal laser scanning microscopy. Ten extracted human teeth showing no clinical signs of caries were investigated. All teeth were horizontally sectioned to approx. 200 m thickness and sections were subjected to different pretreatment conditions as follows vacuum only, ultrasonication only, sodium hypochlorite only, sodium hypochlorite and vacuum, sodium hypochlorite and ultrasonication, and a combination of all three. Surface images were obtained using confocal laser scanning microscopy. Subsurface imaging to a depth of approx. 60 m was achieved using confocal laser microscope techniques. This novel fluorescence technique offers a new alternative for visualization and quantification of dentin.